Sains Malaysiana 45(12)(2016): 1969–1979

http://dx.doi.org/10.17576/jsm-2016-4512-22

 

Perbandingan Filogenetik Protein Antigen-I yang Berpotensi sebagai Calon Diagnostik dan Vaksin terhadap Parasit Cryptocaryon irritans

(Phylogenetic Comparison of I-Antigens Protein as Potential Candidate for Diagnostic

and Vaccine for Cryptocaryon irritans Parasite)

 

Yogeswaran Lokanathan^{1 2}*, Adura Mohd-Adnan^{1 3} & Sheila Nathan¹

 

¹Pusat Pengajian Biosains & Bioteknologi, Fakulti Sains & Teknologi, Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor Darul Ehsan, Malaysia

 

²Pusat Kejuruteraan Tisu, Fakulti Perubatan, Universiti Kebangsaan Malaysia, Jalan Yaacob Latiff

56000 Kuala Lumpur, Wilayah Persekutuan, Malaysia

 

³Fakulti Sains, Teknologi, Kejuruteraan dan Matematik, INTI International University

71800 Bandar Baru Nilai, Negeri Sembilan, Malaysia

 

Received: 14 June 2016/Diterima: 26 October 2016

 

 

ABSTRAK

Protein antigen-i parasit ikan C. irritans berpotensi tinggi digunakan sebagai calon dalam pembangunan vaksin komersial terhadap C. irritans. Walau bagaimanapun, kewujudan variasi pada antigen-i serotip C. irritans yang berbeza mempengaruhi tahap perlindungan yang bakal diberikan terhadap varians C. irritans yang berbeza apabila antigen-i digunakan sebagai vaksin. Kajian ini dijalankan untuk membandingkan jujukan pelbagai antigen-i pencilan C. irritans di Malaysia berbanding antigen-i pencilan C. irritans yang pernah dilaporkan. Perbandingan filogenetik dijalankan untuk meramalkan potensi protein tersebut dalam usaha membangunkan calon serodiagnostik dan pemvaksinan terhadap pencilan C. irritans yang berlainan. Penjajaran jujukan berbilang bagi jujukan asid amino antigen-i dilakukan dengan perisian CLUSTALX dan analisis filogenetik antigen-i dilakukan menggunakan kaedah parsimoni maksimum (MP) dan kaedah Bayes. Sembilan transkrip unik (TU) C. irritans yang mempunyai padanan signifikan dengan antigen-i di pangkalan data protein NCBI didapati mempunyai peratus kesamaan antara 41% hingga 71%. Kedua-dua pohon MP dan Bayesian yang dijana menunjukkan varians antigen-i cn56 and cn57 terkelompok bersama dalam satu kumpulan manakala varians antigen-i yang lain terbahagi kepada dua kumpulan berasingan dan pengkelompokan ini disokong oleh kehadiran asid amino yang terpulihara dalam kumpulan masing-masing. Kajian lanjutan boleh dilakukan untuk mengenal pasti varians antigen-i yang sesuai sebagai calon serodiagnosis dan juga dapat memberi perlindungan silang terhadap pelbagai pencilan C. irritans di serata dunia.

 

Kata kunci: Antigen-i; Cryptocaryon irritans; konformasi; perlindungan silang; serotip

 

ABSTRACT

The fish parasite C. irritans i-antigen protein has high potential to be developed as a commercial vaccine against C. irritans. However, variation within the i-antigens of different C. irritans serotypes affected the cross-protection provided towards different isolates of C. irritans when the i-antigen was evaluated as a candidate vaccine. This study was conducted to compare the sequences of various C. irritans i-antigens isolated in Malaysia with i-antigen sequences of previously reported C. irritans isolates. Phylogenetic comparison was performed to predict the protein’s potential to be developed as a screening marker and vaccine candidate against various C. irritans isolates. Multiple sequence alignment of i-antigen amino acid sequences was undertaken using ClustalX and phylogenetic analysis of i-antigen was performed using the maximum parsimony (MP) and Bayesian methods. Nine C. irritans unique transcripts (UTs) had significant matches with i-antigen sequences in the NCBI protein database, with similarities ranging between 41 and 71%. Both MP and Bayesian trees generated showed that i-antigens cn56 and cn57 variants clustered together in one group while other i-antigen variants were divided over two different groups and the grouping was supported by the presence of conserved amino acids in their respective groups. Further research should identify variants of the i-antigen suitable for serodiagnostic candidates and provision of cross-protection against multiple strains of C. irritans globally.

 

Keywords: Conformation; cross protection; Cryptocaryon irritans; i-antigen; serotype

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*Corresponding author; email: lyoges@ppukm.ukm.edu.my